How to Breed for Behavioral Resistance to Varroa Destructor Krispn Given, Purdue University https://extension.entm.purdue.edu/beehive/
Why breed bees? Increase honey Production Improve overwintering ability Decrease defensive behavior Reduce swarming Reduce or increase use of propolis Change color Decrease incidence of diseases Improve defense against varroa destructor* Other personal reasons
Breeding versus queen rearing
Most important part of a breeding program
Decide on a trait (phenotype) Develop a assay Measure the foundation population Select parents Control mating using a design Evaluate, document, and verify Select parents
Bee Genetics + Breeding Mendel s bee house
Honey Bees 16 CHROMOSOMES ~260 million bases (Mb LOW REPETIVE DNA HAPLOID MALES HIGH RECOMBINATION RATES!
CHROMOSOMES Composed of coiled, twisted DNA and coated with proteins Each species has a specific number They are organized in pairs (one comes from the mother and the other from the father)
Genetic gender and caste Queens lay two types of eggs Males develop from unfertilized eggs Females develop from fertilized eggs
HONEY BEE GENETICS Bees have 16 paired chromosomes. Haplodiploidy Males honey bees have one of each chromosome (haploid 16) Females have two of each chromosome (diploid 32) Polyandry The queen mates with 10 to 15 males, resulting in many sub-families within the hive s family. Many important behaviors are influenced by genes! stinging pollen foraging undertaking hygienic behavior brood rearing traits swarming tendency propolis collection honey production even learning!
FEMALES crossingover X X X
HYBRID CROSS WHAT HAPPENS WHEN YOU CROSS TWO DIFFERENT STOCKS OF BEES or HIGH LINE LOW LINE HYBRIDS X gives you
35% Trota US manag 1ed honey be 1e colonies Loss Estimates rr::&---1 _:5'Mt: 7 - - 1 DAcceptabl e Wintelf Loss D T1olal Winte r Loss T otail Annual loss,... -... _ - -=-! -...,...,_ -=- ' - ' l.5% 5% 0% - - -I I -- - _ I." - - -,_... -... - -,_,.... -... - -! --= - - -- IJ " - i - - - - - - - - ' - >- i- I I -
Registration # Product Name 2724-406 ZOECON RF-318 APISTAN STRIP 2724-406-62042 AP STAN ANTI-VARROA MITE STR PS Active Ingredient Fluvalinate (10.25%) 61671-3 FOR-MITE Formic acid (65.9%) 70950-2 AVACHEM SUCROSE OCTANOATE [40.0%) 70950-2-2205 SUCROCIDE Sucrose octanoate (40%) 70950-2-84710 SUCRASHIELD 73291-1 API LIFE VAR Thymo l (74.09%), Oi l of eucalyptus ( 16%), Menthol (3.73%) 75710-2 MITE-AWAY QUICK STRIPS Formic acid (46.7%) 79671-1 APIGUARD Thymol (25%) 83623-2 HOPGUARD II Hop beta acids resin (16%) 87243-1 Apivar Amitraz (3.33%) 9 1266-1 91266-1-73291 91266-1-91832 OXALIC ACID DIHYDRAT E OXALICACID D HYDRATE OXALICACID D HYDRATE 115 56-138 CHECKMITE+ BEE H IVE PEST Coumaphos ( 10%) CONTROL STRIP 11556-138-61671 CHECKM TE+ BEE HIVE PEST CONTROL STR IP Oxali c acid ( 100%)
Adult female invades cell before its sealed Dr. Harry Laidlaw
female immerses herself in brood food, lays first egg after about 72 hours.
First egg laid is a haploid male, then mites mate in cell and must mature before the bee emerges (usually about 4 females emerge).
Varroa mites prefer drone brood but also invade worker cells. Most mites (80%) will be in the sealed drone and workerbrood
THEY CAME FROM ASIAN HONEY BEES In Apis cerana Varroa only reproduces in drone brood
40k 30k 20k 10k 0 Cell of Bro'od v / Varroa Population Dynamics \ \ / \ / \ I " I / I / ----- I I \ >\ "' r --_ - Mite No. - 0 Apl May Jun Jul Aug Sep Oct Nov Dec 3k 2.5k 2k 1.5k 1k 0.5k Jan Feb Mar
PARASITIC MITE SYNDROME DWV
Virus and brood diseases appear
some vectored viruses from varroa Acute bee paralysis (ABPV) Israel acute bee virus (IAPV) Kashmir bee virus (KBV) Black queen cell virus (BQCV) Deformed wing virus (DWV) 22 known viruses!
Monitoring Mites in Your Hive Uncapping drone brood Ether Roll Ethanol wash Sugar Roll Sticky Boards
Looking for mites in drone brood
Looking for mites on adults
WHAT TRAIT WILL YOU MEASURE AND HOW TO MEASURE IT?
1)How do you choose the parents? (need to evaluate and keep good records) 2)how do you control mating's? (need to do I.I. or have drone source flooding in your mating yard)
PRACTICAL WAY TO SELECT FOR VARROA RESISTANCE:
GRAFT FROM STRONG HIVES THAT DON T GROW MITES
You need try to control mating's Try to have good colonies with drones in the mating yard.
You need a trait you can evaluate, something that is beneficial, and that is heritable. Start with bees that have good traits for your area!
1 1 0.9 0.8 0.7 PROBLEM: Loss of introduced genes open mating for 10 generations. 0.6 0.5 50.00% 0.4 0.3 25.00% 0.2 12.50% 11.25% 0.1 0 5.63% 2.81% 1.41% 0.70% 0.35% 0.18% 1 2 3 4 5 6 7 8 9 10
% DEAD BROOD REMOVED Hygienic Behavior is Heritable 100% 50% 0 (W. Rothenbuler 1964) Generations of Selection (made up data)
Another example: Response to Selection for Pollen Hoarding
USDA Baton Rouge bee lab
VSH BEHAVIOR Varroa-sensitive hygienic behavior (VSH) Greg Hunt Rodger Dewhurst 2006 and 2008 Detection and removal of varroa infested brood University of Warwick www.ces.ncsu.edu
Making uniform colonies
VARROA MITES INSIDE THE CELL
Good age to check for reproductive mites
VSH is an indirect attack on Varroa
extension.org/bee_health
capped cell uncapped Measure changes in: % uncapped % chewed % recapped % varroa infestation % reproductive varroa % varroa with no progeny Visible w/o magnification. but imprecise Precise, but require magnification chew, remove / ' recap Figure 1. - Types of changes in brood combs that can be measured without and with a stereo microscope. For example, the percentage of pupae uncapped during a certain period of t m e could potentially be used to breed for varroa resistance. Uncapped pupae are visible without a microscope, but measuring the % uncapp ed pupae is highly var able and does not always predict VSH behavior. On the other hand, measuring the percentage ofinfertile mites (those without progeny) predicts strong varroa resistance, bu tit req uires detailed microscopic exam inations of brood to measure.
Days past capping Brood stage 3 prepupa Eldest offspr ingin normally 1eproduci ng mites 4 pupa, fi rst,proto white 1eyes nymphe (ma1l1e) 5-6 pupa, second protopnk eyes ny1mph (female} 7-9 pupal purp1e eyes first femare deu1onymph 10-11 pupat, first adult brown headl. black daughter eyes
Uncapping of mite-infested cells due to VSH
24hrs LATER!
GENERAL HYGIENIC BEHAVIOR Freezing capped brood with liquid nitrogen (250 ml) OR PIN PRICK METHOD Greg Hunt
GROOMING BEHAVIOR
MIGUEL AND ERNESTO S STUDY In Mexico Found that grooming behavior was impor tant Those with the lowest infestation overall had low adult infestation show highest mite drop! They also had the most chewed mites on board, and also in the lab assay.
% Mites Chewed in Hives 60 50 40 30 20 10 0 Russians/ Unselected High/Low Gen. 1 High/Low Gen. 2 Africanized/ European
% Mites Removed in Assay 30 25 20 15 10 5 0 Russians/ Unselected High/Low Gen. 1 High/Low Gen. 2 Africanized/ European
What is responsible for behavioral traits? Genotype of queens and workers Environmental factors
Grooming Behavior
MITES IN BROOD % INFESTATION 50 40 30 20 10 0 2 3 4 MONTH 5 6 LOW GROUP HIGH GROUP
MITES ON ADULTS % INFESTATION 35 30 25 20 15 10 5 0 LOW GROUP HIGH GROUP 2 3 4 MONTH 5 6
Selecting fo r c hew ed mite s is stic k y business! '
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Current Selection Program: Grooming and Biting Behavior
Hierarchical Selection: Measure proportion chewed mites. Eliminate those that show disease or did not control mite population growth. Eliminate those that do not pass freezekilled hygienic test. Select breeder colonies.
Started with SURVIVORS, VSH + Russians Method for the first 7 years: Measure mite drop (and strength) multiple times Measure honey yield Test best sources for (VSH) hygienic behavior Use instrumental insemination.
Response to selection at the colony level is also at the individual Initiating grooming dance* grooming Biting VSH General hygiene Undertaker
Instrumental Insemination High x high chewers are crossed and monitored For more response to selection Daughter queens are then grafted from these*
Collecting and counting chewed mites
(p = 0.028)
Photo Mites t h at h ave been chewed by the m ite-b iters ark
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2014 August P<0.001
50 MITE-BITING IS A HERITABLE TRAIT. 45 40 PERCENT CHEWED 35 30 25 20 15 10 5 0 2006 2007 2008 2009 2010 2011 YEAR 2012 2013 2014 2015
Wax moth larva The bite of the bee contains 2-heptanone Varroa
How to select for bees that bite varroa mites and groom them from their bodies Krispn J. Given, Greg J. Hunt, David M. Shenefield, Ginger D. Davidson Dwight C. Wells and Dan P. O Hanlon n Abstract Grooming behavior is one of the key mechanism honey bees (Apis Melifera) execute to kill and remove Varroa destructor from their bodies. Two of the grooming behaviors identified with chewing and mite removal are responsible. Through selection with instrumental insemination we have seen Varroa Destructor chewing levels go from 3% to 43%. The Heartland Honey Bee Selection Process Breeders Cooperative was initiated to promote the production and dissemination of northern mite-resistant stocks. Promoting Breeding Grooming behavior is one of the mechanism of defense against Varroa destructor. Mite grooming behavior is the ability to remove mites and is associated with chewed mites. Figure 1 Chewed mites under l Mites are placed on there idiosoma and inspected for damage. Inaugural HHBBC at the Purdue University bee laboratory, where 75 queens were in distributed. Breeders from PA, WV, OH, attended the three day event. l This shows a a response to our selection. We may be near the peak for grooming behavior but need make sure every colony has a high levels as possible by requeening the low chewers. We are working on a better measure for grooming behavior, and have a better correlation between % chewed mites and reduced mite populations in 2014 than we did in 2013, in 2013 the mitepopulations were higher and more variable between colonies.. Annual queen rearing classes taught at the Purdue bee lab, in 2014 we had 34 students. Instrumental insemination classes have also been a good resource for breeders to learn more about the importance of selection at the Purdue Bee Lab.
2016 Insemination Fest PA, OH, WV, MI, IN, IL, KY, TN 130 queens inseminated
COMMUNITY STOCK EVALUATION IN 2014-2015 54 queens, 23 beekeepers, 3 commercial sources, 2 Purdue sources.
Commercial hives had 3X more mites PHOTO: RATNATHAPA
60% 50% 40% 30% Survival 2014-2015 16/27 = 59 % 20% 10% 6/27 = 22 % 0% Commercial Queens IN Mite Biters
60% 50% 40% 30% 17/27 = 63 % *DIED IN WINTER 2014-2015 20% 10% 7/27 = 26 % 0% Commercial Queens IN Mite Biters
IN QUEENS MADE 40 LB MORE HONEY
Ten of eleven beekeepers preferred IN mite biters
HAS July 13-15, 2017 Evansville, INJuly 13
2016 I.I. Class
Future plans: Cage bee assays for grooming!
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W e lcom e to t h e Hunt lab's B ee Hive. W e are in v o lved in honey bee r esearc h a n d e d ucatio n. J uly 9-11, 201 5 W i s you were },ere C 1 ol - 11- rl aiul A p i cullural,:,od e Y- -.heartlandbees.o rg 14T H A N N UAL H.A.S. CO N FERENCE f ah..ring sesa1ons with : Dennis vanengetsdo<j>, Sue Cob<ty. Larry Connor, JoeTraynor, Clanmce Collison Kim F lottum, Renata Borba, Roger Hoopln11a ner Ken Schramm, ghan Milbrath c o nference s ponsored b y : Bee Culture,, beg to adv. wort.shop,., b<jildjng nucs spl ts, q-ueen rai.sing. drone catch tng. new propotis d scoveries honey tastin g b nng 3 jars to swap! 0 - -- --. -...,./'lo._...... CHILDREN'S PROGRAMMING : cand le - makin g, murals, seed bombs Events Do you have a swa rm to pick up? Find a local swarm catcher here: http:llwww.in.govldnr/entomolols755.htm. When bees swarm the queen and most of the hive bees leave after filling up on honey. Bees in a swarm are usually very gentle but leave them alone! With all those bees flying one could get stuck in your hair.the swann sends out scout bees to find a new cavity to nest in. They usualy stay for a few days and then they are off to their new home. P ur due U ni versit I Colle ge of Agr icul ture I En tomo logy I Extension CoQY...dght 2008, Pu r d ue University, all rights reserved. site author Greg Hunt Website developed by the Entomology Department at P ur due Universit y If you have trouble access in g this page because of a d isability, please contact the Webmaster.
Thank you Heartland Honey Bee Breeders Coop National Honey Board Funding NEW USDA Funding!!! Greg Hunt Jennifer Tsuruda Gladys Andino Jeff Berta - PA Matt Evans IN Tammy Horn - KY Joe Kovalevski OH Scott Martin - IL Megan Milbraith - MI Dan O Hanlon - WV Dave Shenefield - IN Dwight Wells - OH